Isolated cardiomyocytes were loaded with 10 µM Indo-1 AM and electrically stimulated at 1 Hz using a two-platinum electrode insert connected to a bipolar stimulator (SEN-3301; Nihon Kohden) on the stage of an inverted microscope (IX71; Olympus) with a 20× water immersion objective lens (UApo N340; Olympus). Calcium transients were measured as the ratio of fluorescence emitted at 405/480 nm, after excitation at 340 nm, using a high-performance Evolve EMCCD camera (Photometrics). Cardiomyocytes were maintained under continuous flow in standard Tyrode’s solution, exchanged using a microperfusion system. The experiments were recorded and analyzed using MetaMorph software (version 7.7.1.0; Molecular Devices). To estimate the effect of colchicine on cell contraction, we calculated the efficiency of converting elevated Ca2+ levels to cell contraction by dividing the percentage of cell shortening by the Ca2+ amplitude.
Uapo n340
Manufactured by Olympus
The UApo N340 is a high-quality objective lens designed for microscopy applications. It features a numerical aperture of 1.40 and a working distance of 0.13 mm, making it suitable for a variety of imaging techniques. The lens is optimized for use with Olympus microscopes and can provide excellent optical performance.
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Lab products found in correlation
3 protocols using uapo n340
1
Cardiomyocyte Calcium Transient Analysis
2
Calcium Transients in Paced Cardiomyocytes
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Isolated cardiomyocytes were loaded with 10 μmol l−1 Indo-1 AM (Invitrogen) and electrically stimulated at 1 Hz using a two-platinum electrode insert connected to a bipolar stimulator (Nihon Kohden, SEN-3301) on the stage of an inverted microscope (IX71, Olympus) with a × 20 water immersion objective lens (UApo N340, Olympus). Calcium transients were measured as the ratio of fluorescence emitted at 405/480 nm after excitation at 340 nm using a high-performance Evolve EMCCD camera (Photometrics). Cardiomyocytes were maintained under continuous flow in standard Tyrode’s solution, exchanged using a microperfusion system. For measuring caffeine-induced calcium transients, cells were paced at 1 Hz prior to induction of caffeine contractures. Electrical stimulation was stopped 15 s before rapid perfusion with a 10 mmol l−1 caffeine solution. The experiments were recorded and analysed using MetaMorph software (version 7.7.1.0; Molecular Devices). Results were the means of the fluorescent signals from 10–20 cardiomyocytes from a single heart.
3
Electrical Stimulation of Cardiomyocytes
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Isolated cardiomyocytes were electrically stimulated at 0.5, 1, 2, 4 Hz using a two-platinum electrode electrode insert connected to a bipolar stimulator (Nihon Kohden, SEN-3301). Cell images were recorded at a time resolution of 20 ms with a High-performance EvolveTM EMCCD camera (Photometrics) coupled to an inverted microscope (IX71, Olympus) with a 20× water immersion objective lens (UApo N340, Olympus) and analysed using MetaMorph software (version 7.7.1.0; Molecular Devices).
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